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. 1998 Nov;43(5):656–663. doi: 10.1136/gut.43.5.656

Urokinase and the intestinal mucosa: evidence for a role in epithelial cell turnover

P Gibson 1, I Birchall 1, O Rosella 1, V Albert 1, C Finch 1, D Barkla 1, G Young 1
PMCID: PMC1727298  PMID: 9824347

Abstract

Background—The functions of urokinase in intestinal epithelia are unknown. 
Aims—To determine the relation of urokinase expressed by intestinal epithelial cells to their position in the crypt-villus/surface axis and of mucosal urokinase activity to epithelial proliferative kinetics in the distal colon. 
Methods—Urokinase expression was examined immunohistochemically in human intestinal mucosa. Urokinase activity was measured colorimetrically in epithelial cells isolated sequentially from the crypt-villus axis of the rat small intestine. In separate experiments, urokinase activity and epithelial kinetics (measured stathmokinetically) were measured in homogenates of distal colonic mucosa of 14 groups of eight rats fed diets known to alter epithelial turnover. 
Results—From the crypt base, an ascending gradient of expression and activity of urokinase was associated with the epithelial cells. Median mucosal urokinase activities in each of the dietary groups of rats correlated positively with autologous median number of metaphase arrests per crypt (r=0.68; p<0.005) and per 100 crypt cells (r=0.75; p<0.001), but not with crypt column height. 
Conclusions—Localisation of an enzyme capable of leading to digestion of cell substratum in the region where cells are loosely attached to their basement membrane, and the association of its activity with indexes of cell turnover, suggest a role for urokinase in facilitating epithelial cell loss in the intestine. 



Keywords: urokinase; intestinal epithelium; colon; epithelial proliferation

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Figure 1 .

Figure 1

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Immunohistochemical staining of sections of histologically normal colonic mucosa from patients with colorectal cancer. (A) Urokinase staining of upper crypt and surface epithelium (original magnification ×400). Arrows show specific staining of cells with morphology consistent with macrophages. (B) Negative control for (A). (D) and (C) Higher power magnification (original magnification ×800) of colonic sections from two other patients showing staining of cytoplasm of surface colonic epithelial cells, and of the basal and underlying basement membranes. 


Figure 2 .

Figure 2

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Immunohistochemical staining for urokinase of a section of normal human duodenal mucosa. (A) High power magnification (original magnification ×800) of the tip of a duodenal villus showing cytoplasmic staining for urokinase. Enterocytes in the lower villus and crypt showed minimal staining for urokinase (not shown). (B) Negative control of (A). 


Figure 3 .

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Immunohistochemical staining for urokinase in terminal ileum and colon affected by inflammatory bowel disease. (A) Terminal ileum of a patient with Crohn's disease showing cytoplasmic staining of villous but not crypt epithelium (original magnification ×400). (B) Colonic mucosa of a patient with ulcerative colitis, showing distorted crypt architecture but heavy staining of surface epithelial cells and underlying basement membrane (original magnification ×400). 


Figure 4 .

Figure 4

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(A) DNA content, (B) alkaline phosphatase activities (ALP), and (C) urokinase (uPA) activities in enterocyte cell fractions from proximal jejunum, mid-jejunum, and ileum of normal rats. Results are shown as mean (SEM). Statistically significant differences (ANOVA) were found across cell fractions for DNA content (all p<0.001), alkaline phosphatase activities (p<0.001 for proximal and mid-jejunum, p=0.009 for ileum), and urokinase activities for proximal jejunum and ileum only (p<0.001 for both). 


Figure 5 .

Figure 5

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Comparison of mean alkaline phosphatase activities with mean urokinase (uPA) activities across 12 cell fractions from (A) proximal jejunum (r=0.95), (B) mid-jejunum (r=0.99), and (C) ileum (r=0.93). Correlations were all highly significant (p<0.0001; linear regression analysis). 


Figure 6 .

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Correlation of (A) median crypt column height, (B) metaphase index, and (C) number of metaphase arrests per crypt column of distal colonic epithelium with median uPA activities (logarithm transformed) in homogenates of adjacent colonic mucosa in normal rats ingesting a variety of diets. Pearson correlation coefficients were −0.25 for crypt column height (p>0.1), 0.68 (p<0.005) for the number of arrests per crypt, and 0.75 (p<0.001) for the metaphase index. 


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